How difficult is it to extract and crystallize alpha-amylase from human saliva?

In summary: The difficulty rating for crystallizing thermally unstable proteins like alpha amylase would be about a 5.
  • #1
eptheta
65
0
I wanted to perform some experiments on enzyme rates and chose amylase as it is easy to obtain. However, I wanted it in pure form so that I can perform mass/concentration-based calculations as well.
Is there any way to extract alpha-amylase from human saliva?

Thank you.
 
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  • #3
Honestly I have very little equipment. My chem lab is equipped with only basic volumetric tools and random chemicals.
What equipment would I need ? I may be able to access a BSc Chemistry lab, but I am unsure what it does and does not have...
I do know that it has electrophoresis machines and column chromatography facilities.

-No beer making stores near here... Only beer selling.
-My last resort would be buying the amylase enzyme...

This link was pointed out to me.. It's for precipitating out alpha amylase using ethanol. The steps are clear and followable, and the chemicals seem to be easily obtainable, could you check it out and let me know what you think of it ?
http://www.science.smith.edu/departments/Biochem/Biochem_353/amylase.html [Broken]

Thanks !
 
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  • #4
Sounds like you are going to need very pure ethanol, glycogen, a spectrometer, volumetric pipettes and a nice centrifuge. Good luck with it.

The centrifuge isn't something you usually find in the chem lab. Try the biology lab instead. You are going to need a high speed (10,000 g!) one.

I'd prefer to buy the amylase online ($1.95 plus shipping) rather than go through all of this. What the heck is "Oyster glycogen, Sigma, Type II"? Probably something you buy from Sigma-Aldrich.
 
  • #5
chemisttree said:
What the heck is "Oyster glycogen, Sigma, Type II"? Probably something you buy from Sigma-Aldrich.
I concur. Getting Oyster Glycogen is probably way harder than getting amylase...
I guess you're right. I'll just buy the enzyme from here. (Although i think it might be difficult to acquire it in Mumbai, India)

One more thing, on a scale of 1 to 10, how difficult do you think it is to crystallize thermally unstable proteins like alpha amylase ? (Hoping that you have experience with this kind of thing)
I wanted to crystallize it for fun, but i realize that its nothing like crystallizing rochelle's salt or sucrose (both of which require lots of heating to make a supersaturate)

Thanks!
 

What is alpha amylase extraction?

Alpha amylase extraction is a process in which the enzyme alpha amylase is isolated and purified from a natural source such as plants, animals, or microorganisms. This enzyme is responsible for breaking down carbohydrates into smaller molecules, and is commonly used in various industries such as food and beverage, pharmaceutical, and textile.

What are some common sources of alpha amylase for extraction?

Alpha amylase can be extracted from various sources including barley, wheat, rice, corn, potatoes, and certain bacteria and fungi. These sources contain high concentrations of the enzyme, making them ideal for extraction.

What is the purpose of alpha amylase extraction in the food industry?

In the food industry, alpha amylase is used to break down starch into simple sugars, which improves the texture and flavor of food products. It is commonly used in bread making, brewing, and production of sweeteners and syrups. It can also be used to reduce the viscosity of sauces and improve the texture of dairy products.

What is the process of alpha amylase extraction?

The process of alpha amylase extraction involves several steps including cell disruption to release the enzyme, filtration to remove solid particles, and purification through techniques such as chromatography. The extracted enzyme is then concentrated and dried to form a powder for commercial use.

What are the benefits of using extracted alpha amylase compared to natural sources?

Using extracted alpha amylase has several benefits compared to using natural sources. It allows for a more controlled and consistent enzyme activity, resulting in better quality and yield of products. It also eliminates the risk of contamination from other enzymes and microorganisms present in natural sources.

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